CN102599079B - Method for constructing half-sib family of Megalobrama Terminalis - Google Patents
Method for constructing half-sib family of Megalobrama Terminalis Download PDFInfo
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- CN102599079B CN102599079B CN201110028552XA CN201110028552A CN102599079B CN 102599079 B CN102599079 B CN 102599079B CN 201110028552X A CN201110028552X A CN 201110028552XA CN 201110028552 A CN201110028552 A CN 201110028552A CN 102599079 B CN102599079 B CN 102599079B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention belongs to the technical field of fresh water fish breeding, and specially relates to a method for constructing half-sib family of Megalobrama Terminalis. The method comprises (1) selecting Megalobrama Terminalis with body weight larger than 0.5 Kg, labeling with electronic label, and performing individual culture in advance for 6 months; (2) in June of current year, regulating sex gland development to reach synchronism, and performing primary injection of human chorionic gonadotropin (HCG) on female fish; (3) performing secondary injection of HCG and domperidone (DOM) on female and male parents 12 hr after the primary injection; (4) constructing half-sib family of Megalobrama Terminalis: one female parent crossed with two male parents for maternal half-sib family; one male parent crossed with two female parents for patrilineal half-sib family; (5) after the secondary injection, if ova automatically flow out when slightly pressing abdomen of Megalobrama Terminalis with finger, performing artificial insemination; (6) dividing collected ova into two equal parts, placing in stainless steel bowls, inseminating with sperm of two male fishes, debonding with loess water, rinsing with clear water, and placing in hatching jar.
Description
Technical field
The invention belongs to fish breeding method field, be specifically related to a kind of construction method of megalobrama amblycephala family half sibs.
Background technology
Megalobrama amblycephala (Megalobrama amblycephala Yih, 1955), have another name called blunt snout bream, is under the jurisdiction of Osteichthyes, Cypriniformes, and Cyprinidae, triangular bream belongs to, and its natural distribution only is confined in a few lake, the middle and lower reach of Yangtze River.Megalobrama amblycephala is one of important phytophagy economic fish of China, because its feeding habits are wide, cost is low, growth is fast, survival rate is high, easily fish for, can be in pond egg reproduction, all have delicious, little, contain the advantages such as the meat rate is high, the bodily form good, specification is moderate, thereby be used as good herbivorous fishes kind and generally promote in the whole nation.
In recent years, megalobrama amblycephala becomes the market best selling variety, and fanning economics is better.Thereby market is very urgent to the demand of the good seed of megalobrama amblycephala, the seed breeding production of megalobrama amblycephala urgently to be speculated, plasm resource protection and the fine-variety breeding process of quickening megalobrama amblycephala.Due to the inbred phenomenon of ubiquity in the production of megalobrama amblycephala seed, cause germplasm to be degenerated and aggravate.The growth rate of conventional seed is slower, has not only affected the growth performance of megalobrama amblycephala, and has restricted to a certain extent megalobrama amblycephala in wider popularization.Although China has selected growth rate megalobrama amblycephala new varieties " No. one, Pujiang " faster, its selection adopts traditional colony's selection-breeding method, its offspring's Character instability.At present domestic urgently the excavation and the Innovation Germplasm resource, select the megalobrama amblycephala improved seeds of fast growth, good stress resistance.
In the seed selection of aquatic livestock, more and more be inclined to and use extensive family selective breeding technology in the world at present, and use extensive family selective breeding, build family half sibs and be absolutely necessary, because in seed selection, a lot of genetic parameters all will be estimated by family half sibs.There is report to think, at rainbow trout (Kincaid et al.1977; Gjerde 1986), Atlantic salmon (Gjerde 1986,1999), Tilapia mossambica (Longalong et al.1999; Gall et al.2002), channel catfish (Rezk et al.2003), silver salmon (Cerda et al, 2003), Penaeus Vannmei (Fjalestad et al.1997; Hetzel et al.2000), east oyster (Davis 2000) and domestic to Chinese prawn (field spark 2007; Yang Cui China 2007) existing successful implementation case and in the seed selection of turbot (Luan Sheng etc. 2008), long oyster (Wang Qingzhi etc., 2009) etc.Yet, in the family half sibs of actual implementation megalobrama amblycephala, still exist the parent to grow asynchronous, the quality and quantity that raun lays eggs is unstable, milter seminal fluid deficiency, causes half sibs to build the not high outstanding problem of efficiency.And, because parent's growth is inconsistent, making half sibs build asynchronous, construction schedule is also long.
There is not yet up to now the relevant report of the construction method of relevant megalobrama amblycephala family half sibs.
Summary of the invention
The object of the invention is to overcome the defect of prior art, a kind of construction method of megalobrama amblycephala family half sibs is provided, the method can meet follow-up extensive megalobrama amblycephala family selective breeding requirement.
The present invention is achieved through the following technical solutions:
A kind of method that builds the megalobrama amblycephala family half sibs, its step comprises:
1) in nature early January in time, select the megalobrama amblycephala that 2 ages, above and body weight was greater than 0.5kg to carry out in advance individuality cultivation, with the PIT electronic marker, be injected into megalobrama amblycephala dorsal fin base portion by syringe, carry out respective record, cultivated 6 months;
2) megalobrama amblycephala in June then, harvesting step 1), manual injection's hormone is regulated its gonad development makes it synchronous, while injecting hormone for the first time, only injects raun, and the hormone of injection is human chorionic gonadtropin, and injected dose is 1 μ g/kg body weight;
3) after injecting for the first time hormone 12h, female, male megalobrama amblycephala is injected to hormone for the second time, the hormone of injection and dosage human chorionic gonadtropin are 4 μ g/kg body weight; DOM is the 2mg/kg body weight;
4) build the megalobrama amblycephala family half sibs: wherein build maternal family half sibs by the female male proportional arrangement of 2 tails of joining of megalobrama amblycephala 1 tail, or the fish brood amount of female megalobrama amblycephala builds the maternal family half sibs of megalobrama amblycephala over 500,000 in the female male ratio of 3 tails of joining of 1 tail; Wherein build the paternal half-sib family female by male 2 tails of joining of megalobrama amblycephala 1 tail, or individual sperm volume is over 2ml, by the male female structure paternal half-sib of the 3 tail family of joining of 1 tail;
5) in step 3) after megalobrama amblycephala injects hormone 6-8h for the second time, press female megalobrama amblycephala belly ovum automatically to flow out with light finger, can carry out artificial insemination;
6) the megalobrama amblycephala raun is pulled out, wiped body surface moisture, ovum is clamp-oned in the stainless steel bowl of drying, the ovum of getting is divided into to 2 deciles, in two the stainless steel bowls that are placed in, stand-by; Maybe the ovum over 500,000 is classified in three categories minute, in three the stainless steel bowls that are placed in, stand-by; Then in the described stainless steel bowl split, clamp-on the seminal fluid (about 0.8-1.1ml) of the male megalobrama amblycephala of 2 tail, then add 20ml physiological saline, be stirred to gently 3-4min; With yellow mud water, the fertilized egg of megalobrama amblycephala is carried out to unsticking respectively, clear water rinses the hatching of rear placement hatching cylinder and gets final product;
Wherein
Step 2) step of described individual cultivation is: cultivation separately, breeding density is every 666.7m
2Water surface for aquaculture is put the 200-300 tail in a suitable place to breed, and during cultivation, the proportioning of feed diet is that 30% grass carp is raised together with the buoyancy commercial feed, 70% green fodder (such as perennial ryegrass or Sudan grass etc.) by weight.
Step 3) and 5) determine according to the water temperature situation at the interval of the time of double injection hormone, and when water temperature was 20-22 ℃, be 12h interval time; During higher than 22-25 ℃, be 8h interval time when water temperature.
Step 6) described physiological saline, be the physiological saline of 0.6% concentration, is about to 0.6g sodium chloride and adds a small amount of distilled water and dissolve and be settled to 100ml.
Step 6) described yellow mud water is that yellow mud adds water and fully stirs evenly rear mistake 40 order nets and get final product.
The present invention has the following advantages:
1. the present invention is not high to equipment requirement, easy and simple to handle, and the megalobrama amblycephala family half sibs is built into power and significantly improves, and in common laboratory and cultivation place all can reach, and structure megalobrama amblycephala family also only needs 3-4 people can carry out the batch structure.
2. quantity that can each family of balance when building the megalobrama amblycephala family, the uniformity with the offspring that guarantees each family, efficiently solve family half sibs and build nonsynchronous problem, greatly shortened the cycle that family builds, and is easy to carry out family standardization cultivation.
3. the present invention is little to the damage of megalobrama amblycephala parent population, and the parent can continue on for breeding in the coming year, and the filial generation survival rate is high.
Embodiment
Application describes to the invention process below in conjunction with embodiment.
Embodiments of the invention are to implement at national megalobrama amblycephala seed farm (Ezhou, Hubei Province), the megalobrama amblycephala parent population of selecting come to Jiangxi Poyang Lake, Hubei Province's Liangzi Lake and and the wild megalobrama amblycephala of the Yuni Lake in Hubei Province, totally 73 tails, female 24 tails wherein, male 49 tails.
Test in then about December, will cultivate separately be used to the parent who builds the megalobrama amblycephala family half sibs, namely breeding density is 200kg/666.7m
2Water surface for aquaculture.By weight, the daily ration ratio of feeding is that the green grass (as perennial ryegrass and the Sudan grass of plantation) that 30% grass carp is raised together with buoyancy commercial feed (purchased from Wuhan Gao Long feed corporation,Ltd) and 70% is fattened, before being continued until the Second Year breeding in June.
About June in next year, when temperature of cultivation reaches 20-22 ℃, the megalobrama amblycephala parent population has been caught, with PIT electronic marker (rising barcode technology Co., Ltd purchased from the Guangzhou flood), be injected into megalobrama amblycephala dorsal fin base portion by syringe, the male and female megalobrama amblycephala is separately supported and fasting temporarily, at storage pond, holding pond, give sufficient aeration and be greater than 5mg/L to guarantee dissolved oxygen.
Between table 1 megalobrama amblycephala in age in June family half sibs body weight and body long comparison
By the double injection hormone, carry out genital regulating and guarantee that parent's gonad development is synchronous.Concrete steps are as follows:
1) in June then, the megalobrama amblycephala parent that harvesting is cultivated, manual injection's hormone is regulated its gonad development makes it synchronous, while injecting (18:00-20:00 carries out) hormone for the first time, only inject raun, the hormone of injection is human chorionic gonadtropin, and injected dose is 1 μ g/kg body weight;
2) after injecting for the first time hormone 12h, female, male megalobrama amblycephala is injected to (6:00-8:00 carried out on 1) hormone for the second time, the hormone of injection and dosage human chorionic gonadtropin are 4 μ g/kg body weight; DOM is the 2mg/kg body weight;
3) build the megalobrama amblycephala family half sibs: wherein build maternal family half sibs by the female male proportional arrangement of 2 tails of joining of megalobrama amblycephala 1 tail, or the fish brood amount of female megalobrama amblycephala builds maternal family half sibs over 500,000 in the female male ratio of 3 tails of joining of 1 tail; Wherein build the paternal half-sib family female by male 2 tails of joining of megalobrama amblycephala 1 tail, or individual sperm volume is over 2ml, by the male female structure megalobrama amblycephala of the 3 tail paternal half-sib family of joining of 1 tail;
4) in step 2) after megalobrama amblycephala injects hormone 6-8h for the second time, press megalobrama amblycephala belly ovum automatically to flow out with light finger, can carry out artificial insemination;
5) the megalobrama amblycephala raun is pulled out, wiped body surface moisture, ovum is clamp-oned in the stainless steel bowl of drying, the ovum of getting is divided into to 2 deciles, in two the stainless steel bowls that are placed in, stand-by; Maybe the ovum over 500,000 is classified in three categories minute, in three the stainless steel bowls that are placed in, stand-by; Then to the seminal fluid 0.8-1.1ml that clamp-ons megalobrama amblycephala 2 tail milters in the described stainless steel bowl split, add 20ml physiological saline, be stirred to gently 3-4min; Used respectively the yellow mud water of 40 order nets to carry out unsticking to the fertilized egg of megalobrama amblycephala, and until fertilized egg is not agglomerating, then rinsed in rear placement hatching cylinder and carry out Lotic hatching with clear water, hatching density is 2-5 grain/ml.
Determine according to the water temperature situation at the interval of the time of above-mentioned double injection hormone, and when water temperature was 20-22 ℃, be 12h interval time; During higher than 22-25 ℃, be 8h interval time when water temperature.
In when pairing, if parent's gonad development not mature enough (or growing overdone) occurs, select in time the male or female megalobrama amblycephala of a tail to match and remedy.The present embodiment has finally successfully been set up the maternal family half sibs of 18 megalobrama amblycephalas, 12 megalobrama amblycephala paternal half-sib familys.
The present invention has set up altogether 60 family full-sibses by three day time, and all family half sibses successfully construct.With reference to same method, 9 maternal family half sibses have been set up.Building megalobrama amblycephala family half sibs success rate is 100%, and each family filial generation survival rate is all more than 90%.In each family of megalobrama amblycephala that the present invention builds, neat and consistent is grown in filial generation, and effect is in Table 1.
List of references:
Luan Sheng, Kong Jie, Wang Qing prints the progress of .2008. aquatic livestock breeding value method of estimation and application thereof. marine fishery research, 29 (3): 101-107.
Tian Yan, Kong Jie, Luan Sheng, while opening sky, Luo Kun, Wang Qing print the estimation of .2008. Chinese prawn growth traits genetic parameter. marine fishery research .29 (3): 1-6.
Wang Qingzhi, Li Qi, Liu Shikai, Kong Lingfeng, Zheng Xiaodong, in heritability and the correlation analysis thereof of the long oyster larval growth of auspicious extra large .2009. proterties. Chinese aquatic science, 16 (5): 736-743.
Yang Cuihua, Kong Jie, Wang Qingyin, the field Yi, Luo Kun, while opening sky, estimated heritability and the genetic correlation of 6 Immune interrelation components of the intelligent .2007. Chinese prawn of Liu Qing. Science Bulletin: 52:183-186.
Cerda，C.，Neira，R.and?Barria，N.2003.genetic?and?phenotypic?parameters?for?harvest?weight，color?flesh?and?fat?content?in?coho?salmon(Oncorhynchus?kisutch).International?associationfor?genetics?in?Aquaculture，genetics?in?Aquaculture?VIII，9-15?November，2003：102，Puerto?Varas，Chile.
Davis，C.V.2000.Estimation?of?narrow-sense?heritability?for?larval?and?juvenile?growthtraits?in?selected?and?unselected?sub-lines?of?eastern?oysters，Crassostrea?virginica.Journal?of?shellfish?research，19：613.
Fjalestad，K.T.，Gjedrem，T.，CARR，W.H.and?Sweeney，J.N.1997.Final?report：the?shrimpbreeding?program?selective?breeding?of?Penaeus?Vannamei.AKVAFORSK.Report?no.17/97，85pp.
Gall，G.A.E.AND?Bakar，Y.2002.Application?of?mixed-model?techniques?to?fish?breedimprovement：analysis?of?breeding-value?selection?to?increase?98-day?body?weight?in?tilapia.Aquaculture，212：93-113.
Gjerde，B.and?Korsvpll，A.1999.Realized?selection?differetials?for?growth?rate?and?earlysexual?maturity?in?Atlantic?salmon.Abstract，Aquaculture?Europe?99，Trondheim，Norway，August7-10：73-74.
Gjerde，B.1986.Growth?and?reproduction?in?fish?and?shellfish.Aquaculture，57：37-55.
Hetzel，D.J.，S.，Crocos，P.J.，Davis，G.P.，Moorc，S.S.and?Preston，N.C.2000.Responseto?selection?and?heritability?for?growth?in?the?Kuruma?prawn，penacus?japanicus，Aquaulture，181：215-223.
Kincaid，H.L.，Bridges，W.R.and?Von?Limbach，B.1977.Three?generation?of?selection?for?growthrate?in?fall?spawning?rainbow?trout.Trans.Am.Fish.Soc.，106：621-629.
Longalong，F.M.，Eknath，A.E.and?Bentsen，H.B.1999.Response?to?bi-directional?selectionfor?frequency?of?early?maturing?females?in?Nile?tilapia(Oreochromis?niloticus).Aquculture，178：13-25
Rezk，M.A.，Smitherman，R.O.，Williams，J.C.，Nichols，A.，Kucuktas，H.and?Dunham，R.A.2003.Response?to?thee?generations?of?selection?for?increased?body?weight?in?channel?catfish，ictalurespunctatus，grown?in?carthen?ponds.Aquaculture，228，69-79.
Claims (2)
1. method that builds the megalobrama amblycephala family half sibs is characterized in that the following step:
1) in nature early January in time, select the megalobrama amblycephala that 2 ages, above and body weight was greater than 0.5kg to carry out in advance individuality cultivation, with the PIT electronic marker, be injected into megalobrama amblycephala dorsal fin base portion by syringe, carry out respective record, cultivated 6 months;
2) megalobrama amblycephala in June then, harvesting step 1), manual injection's hormone is regulated its gonad development makes it synchronous, while injecting hormone for the first time, only injects raun, and the hormone of injection is human chorionic gonadtropin, and injected dose is 1 μ g/kg body weight;
3) after injecting for the first time hormone 12h, female, male megalobrama amblycephala is injected to hormone for the second time, the hormone of injection and dosage human chorionic gonadtropin are 4 μ g/kg body weight; DOM is the 2mg/kg body weight;
4) build the megalobrama amblycephala family half sibs: wherein build maternal family half sibs and join the male proportional arrangement of 2 tails by megalobrama amblycephala 1 tail is female, or the fish brood amount of female megalobrama amblycephala builds maternal family half sibs over 500,000 in the female male ratio of 3 tails of joining of 1 tail; Wherein build the paternal half-sib family female by male 2 tails of joining of megalobrama amblycephala 1 tail, or individual sperm volume is over 2ml, by the male female structure megalobrama amblycephala of the 3 tail paternal half-sib family of joining of 1 tail;
5) in step 3) after megalobrama amblycephala injects hormone 6～8h for the second time, press megalobrama amblycephala belly ovum automatically to flow out with light finger, can carry out artificial insemination;
6) the megalobrama amblycephala raun is pulled out, wiped body surface moisture, ovum is clamp-oned in the stainless steel bowl of drying, the ovum of getting is divided into to 2 deciles, in two the stainless steel bowls that are placed in, stand-by; Maybe the ovum over 500,000 is classified in three categories minute, in three the stainless steel bowls that are placed in, stand-by; Then to the seminal fluid 0.8-1.1ml that clamp-ons megalobrama amblycephala 2 tail milters in the described stainless steel bowl split, add 20ml physiological saline, be stirred to gently 3～4min; Used respectively the yellow mud water of 40 order nets to carry out unsticking to the fertilized egg of megalobrama amblycephala, clear water rinses rear placement hatching cylinder and gets final product;
Wherein
Step 1) step of described individual cultivation is: cultivation separately, breeding density is every 666.7m
2Put 200～300 tails in a suitable place to breed, during cultivation, the proportioning of feed diet is that 30% grass carp is raised together with the buoyancy feed, 70% green fodder by weight.
2. method according to claim 1, is characterized in that step 3) and 5) interval of time of double injection hormone determines according to the water temperature situation, when water temperature was 20 ℃-22 ℃, be 12h interval time; Higher than 22 ℃ and during lower than 25 ℃, be 8h interval time when water temperature.
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| CN105961251B (en) * | 2016-05-25 | 2018-11-06 | 上海海洋大学 | A kind of construction method of lower oxygen concentration resistance megalobrama amblycephala |
| CN114375881A (en) * | 2022-01-24 | 2022-04-22 | 佛山市南海百容水产良种有限公司 | Efficient construction method for half-sib families of micropterus salmoides |
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